Polynucleotide agents targeting complement component c5 and methods of use thereof

Abstract

The invention relates to polynucleotide agents targeting the complement component C5 gene, and methods of using such polynucleotide agents to inhibit expression of C5 and to treat subjects having a complement component C5-associated disease, e.g., paroxysmal nocturnal hemoglobinuria.

1 . A single-stranded antisense polynucleotide agent for inhibiting expression of complement component C5, wherein the agent is 10-40 contiguous nucleotides in length, wherein at least one of the contiguous nucleotides is a modified nucleotide, and wherein the nucleotide sequence of the agent is at least 80% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO:1, wherein substantially all of the nucleotides of the single-stranded antisense polynucleotide agent are modified nucleotides. 
     
     
         2 . The agent of  claim 1 , wherein the equivalent region is one of the target regions of SEQ ID NO:1 provided in Table 3. 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . The agent of  claim 1 , wherein the agent is 10 to 30 nucleotides in length; 18 to 30 nucleotides in length; 10 to 24 nucleotides in length: 18 to 24 nucleotides in length; or 20 nucleotides in length. 
     
     
         6 . The agent of  claim 1 , wherein the modified nucleotide comprises a modified sugar moiety selected from the group consisting of: a 2′-O-methoxyethyl modified sugar moiety, a 2′-methoxy modified sugar moiety, a 2′-O-alkyl modified sugar moiety, a bicyclic sugar moiety. 
     
     
         7 . The agent of  claim 1 , wherein the modified nucleotide is a 5-methylcytosine. 
     
     
         8 . The agent of  claim 1 , wherein the modified nucleotide comprises a modified internucleoside linkage. 
     
     
         9 . The agent of  claim 1 , comprising a plurality of 2′-deoxynucleotides forming a gap segment flanked on each side by at least one nucleotide having a modified sugar moiety forming a 5′ wing segment and a 3′ wing segment. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . The agent of  claim 9 , wherein the gap segment is 5 to 14 nucleotides in length. 
     
     
         13 . The agent of  claim 1 , wherein the agent further comprises a ligand. 
     
     
         14 . The agent of  claim 13 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative. 
     
     
         15 . A pharmaceutical composition for inhibiting expression of a complement component C5 gene comprising the agent of  claim 1 . 
     
     
         16 . A pharmaceutical composition comprising the agent of  claim 1 , and a lipid formulation. 
     
     
         17 . A method of inhibiting complement component C5 expression in a cell, the method comprising:
 (a) contacting the cell with the agent of  claim 1 ; and   (b) maintaining the cell produced in step (a) for a time sufficient to obtain antisense inhibition of a complement component C5 gene, thereby inhibiting expression of the complement component C5 gene in the cell.   
     
     
         18 . The method of  claim 17 , wherein the cell is within a subject. 
     
     
         19 . The method of  claim 18 , wherein the subject is a human. 
     
     
         20 . A method of treating a subject having a disease or disorder that would benefit from reduction in complement component C5 expression, the method comprising administering to the subject a therapeutically effective amount of the agent of  claim 1 , thereby treating the subject. 
     
     
         21 . A method of preventing at least one symptom in a subject having a disease or disorder that would benefit from reduction in complement component C5 expression, the method comprising administering to the subject a prophylactically effective amount of the agent of  claim 1 , thereby preventing at least one symptom in the subject having a disorder that would benefit from reduction in C5 expression. 
     
     
         22 - 25 . (canceled) 
     
     
         26 . The agent of  claim 9 , wherein the 5′-wing segment is 1 to 6 nucleotides in length. 
     
     
         27 . The agent of  claim 9 , wherein the 3′-wing segment is 1 to 6 nucleotides in length. 
     
     
         28 . The agent of  claim 13 , wherein the antisense polynucleotide agent is conjugated to the ligand at the 3′-terminus.